Tissue specific hypoxia regulated therapeutic constructs

Abstract

Methods and compositions relating to chimeric genes containing (i) a tissue-specific promoter and (ii) a hypoxia response enhancer element, both of which are operably linked to a selected gene, such as a reporter gene, therapeutic gene (e.g., bcl-2, NOS, catalase and SOD), or deleterious gene are disclosed. Expression of the selected gene is enhanced in the target tissue under hypoxic conditions, such as conditions encountered during episodes of ischemia and reperfusion. The methods and compositions may be used as therapeutics and/or diagnostics.

It is claimed: 
     
       1. A chimeric DNA construct, comprising a hypoxia response enhancer element, a tissue-specific promoter heterologous to the element, and a coding sequence, with the proviso that the coding sequence is not an alpha globin coding sequence,   wherein said promoter is operably linked to said coding sequence and said element is effective to cause expression of said coding sequence under conditions of hypoxia.   
     
     
       2. A chimeric DNA construct of claim 1, wherein said promoter is a cardiac-specific promoter. 
     
     
       3. A chimeric DNA construct of claim 1, wherein said promoter is selected from the group consisting of α-MHC 5 .5 promoter, α-MHC 87  promoter, and human cardiac actin promoter. 
     
     
       4. A chimeric DNA construct of claim 1, wherein said promoter is a kidney-specific promoter. 
     
     
       5. A chimeric DNA construct of claim 4, wherein said promoter is a renin promoter. 
     
     
       6. A chimeric DNA construct of claim 1, wherein said promoter is a brain-specific promoter. 
     
     
       7. A chimeric DNA construct of claim 6, wherein said promoter is selected from the group consisting of aldolase C promoter, and tyrosine hydroxylase promoter. 
     
     
       8. A chimeric DNA construct of claim 1, wherein said promoter is a vascular endothelium-specific promoter. 
     
     
       9. A chimeric DNA construct of claim 8, wherein said promoter is selected from the group consisting of Et-1 promoter and vonWillebrand factor promoter. 
     
     
       10. A chimeric DNA construct of claim 1, wherein said hypoxia response enhancer element is selected from the group consisting of erythropoietin HRE element (HREE1), pyruvate kinase (PKM) HRE element, enolase 3 (ENO3) HRE element and endothelin-1 (ET-1) HRE element. 
     
     
       11. A chimeric DNA construct of claim 1, wherein said coding sequence is selected from the group consisting of nitric oxide synthase (NOS), Bcl-2, superoxide dismutase (SOD), and catalase. 
     
     
       12. An expression vector, comprising a hypoxia response enhancer element, a tissue-specific promoter heterologous to the element, and a coding sequence, with the proviso that the coding sequence is not an alpha globin coding sequence   wherein said promoter is operably linked to said coding sequence and said element is effective to cause expression of said coding sequence.   
     
     
       13. An expression vector of claim 12, wherein said expression vector is a plasmid. 
     
     
       14. An expression vector of claim 12, wherein said expression vector is an adenovirus vector. 
     
     
       15. An expression vector of claim 12, wherein said expression vector is a retrovirus vector. 
     
     
       16. The chimeric DNA construct of claim 1, wherein said coding sequence is a viral thymidine kinase coding sequence. 
     
     
       17. The chimeric DNA construct of claim 16, wherein said viral thymidine kinase coding sequence encodes herpes simplex viral thymidine kinase. 
     
     
       18. The chimeric DNA construct of claim 1, wherein said coding sequence encodes luciferase.